Teaching Hardy-Weinberg Equilibrium using Population-Level Punnett Squares: Facilitating Calculation for Students with Math Anxiety.

Hardy-Weinberg (HW) equilibrium and its accompanying equations are widely taught in introductory biology courses, but high math anxiety and low math proficiency have been suggested as two barriers to student success. Population-level Punnett squares have been presented as a potential tool for HW equilibrium, but actual data from classrooms have not yet validated their use. We used a quasi-experimental design to test the effectiveness of Punnett squares over 2 days of instruction in an introductory biology course. After 1 day of instruction, students who used Punnett squares outperformed those who learned the equations. After learning both methods, high math anxiety was predictive of Punnett square use, but only for students who learned equations first. Using Punnett squares also predicted increased calculation proficiency for high-anxiety students. Thus, teaching population Punnett squares as a calculation aid is likely to trigger less math anxiety and help level the playing field for students with high math anxiety. Learning Punnett squares before the equations was predictive of correct derivation of equations for a three-allele system. Thus, regardless of math anxiety, using Punnett squares before learning the equations seems to increase student understanding of equation derivation, enabling them to derive more complex equations on their own.

Female In-Class Participation and Performance Increase with More Female Peers and/or a Female Instructor in Life Sciences Courses.

As we strive to make science education more inclusive, more research is needed to fully understand gender gaps in academic performance and in-class participation in the life sciences. Studies suggest that male voices dominate introductory biology courses, but no studies have been done on upper-level courses. Results on achievement gender gaps in biology vary and often conflict, and no studies have been done on the correlation between participation and academic performance gaps. We observed 34 life sciences courses at all levels at a large private university. Overall, males were more likely to participate than their female peers, but these gender gaps varied from class to class. Females participated more in classes in which the instructor called on most hands that were raised or in classes with more females in attendance. Performance gender gaps also varied by classroom, but female final course grades were as much as 0.2 SD higher in classes with a female instructor and/or a female student majority. Gender gaps in participation and final course grades were positively correlated, but this could be solely because female students are more likely to both participate more and earn higher grades in classes with many females in attendance.

Determination of antimony and barium in UK air quality samples as indicators of non-exhaust traffic emissions.

The aim of this study was to determine concentrations of antimony and barium in air quality samples and assess the potential suitability of these metals as tracer elements for non-exhaust traffic emissions sources. Air quality filters sampled at monitoring sites around the UK for the purposes of the UK Heavy Metals Monitoring Network were subjected to microwave digestion in hydrogen peroxide and nitric acid, then analysed by ICP-MS for a suite of metals including antimony and barium. The average antimony concentration found across all the network sites was 1.84 ng m-3; the average barium concentration was 6.33 ng m-3. The range of antimony concentrations observed was 0.13-8.02 ng m-3; barium concentrations ranged from levels below the detection limit of 0.18 to 39.9 ng m-3. There are no legislative limits for antimony and barium in ambient air, but the maximum concentrations found are well below the Workplace Exposure Limits specified by the UK Health and Safety Executive. The highest concentrations were found at roadside sites situated to monitor traffic environments, supporting the suitability of antimony and barium to be considered tracer elements for traffic emissions sources. Strong correlations were observed between antimony, barium and copper, indicating they share a common traffic-related source. Based on the strong correlation with copper at urban and traffic locations, indicative annual UK atmospheric emission estimates for antimony and barium in brake and tyre wear were calculated as 6 and 19 tonnes respectively.

Neural transplantation of human MSC and NT2 cells in the twitcher mouse model.

BACKGROUND: Accumulating evidence has demonstrated that the NT2 embryonal carcinoma cell line and multipotential stem cells found in BM, mesenchymal stromal cells (MSC), have the ability to differentiate into a wide variety of cell types. This study was designed to explore the efficacy of these two human stem cell types as a graft source for the treatment of demyelinating disorders such as Krabbe's disease and multiple sclerosis (MS). METHODS: We examined the engraftment and in vivo differentiation of adult MSC and NT2 cells after transplantation into two demyelinating environments, the neonatal and postnatal twitcher mouse brain. RESULTS: Both types of xenografts led to anatomical integration, without tumor formation, and remained viable in the normal and twitcher mouse brain, showing differentiation into neurons, astrocytes and oligodendrocytes. DISCUSSION: This study represents a platform for further stem cell transplantation studies in the twitcher model and potentially has important therapeutic implications.

A numerical approach to resonance frequency analysis for the investigation of oral implant osseointegration.

Experimental devices based on vibration testing are employed as non-destructive procedures for evaluating implants osseointegration. Their behaviour was evaluated considering the outcome of numerical analysis. The purpose was to use the finite element method for assessing the ability of frequency analysis in detecting the degree of oral implant osseointegration. A three-dimensional model of a mandible was obtained from tomographic survey. A single implant was considered in canine region. Two configurations were analysed, with and without a mass linked to the implant as a cantilever, reproducing experimental devices. Simulation consisted of analysing the response to impulse forces for different osseointegration levels, thus evaluating the biomechanical efficiency of the implant-bone compound. A good correlation between frequency response and osseointegration level was obtained. This was carried out by providing an impulse excitation of the implant that resulted in a vibration pattern. Within the limit of finite element analysis, the outcomes showed that numerical investigation provides understanding the behaviour of testing devices based on frequency measurements, confirming the potential of vibrations technique as non-invasive analysis for osseointegration process.

The Son-Rise Program intervention for autism: prerequisites for evaluation.

With increasing availability of interventions for children with autism, it is important that these are backed by rigorous evaluation data that have high levels of ecological validity. To achieve this, a key prerequisite for any evaluation is to gather data on typical consumers and typical implementation patterns of the intervention. This study collected such data longitudinally in relation to the Son-Rise Program, a home-based parent-run intervention for autism. Questionnaires and interview data on family demographics, implementation patterns, and perceived treatment fidelity were gathered three times over the course of a year from families who had attended a Son-Rise initial training course. Although it proved possible to produce a profile of intervention use, findings indicated that the programme is not always implemented as it is typically described in the literature. The study also highlighted methodological challenges likely to be encountered in any future evaluation of this and similar interventions for autism.

The effect of feedback training on lumbosacral compression during simulated occupational lifting.

This study measured the effect of a feedback training program on lumbar compression during simulated occupational lifting. Two distinct types of feedback were compared: real-time electromyographic feedback, vs. an acceleration index delivered verbally post-lift. Kinematic data were collected from 28 subjects during symmetrical sagittal plane lifts. Following a baseline session, two feedback training sessions were provided, with a 1-week interval between each session. A control group followed the same protocols, but without receiving feedback training. A post-training session, using protocols identical to the baseline session, was used to assess pre-to-post changes in the dependent variable: peak lumbosacral compression computed using a dynamic linked-segment model. All three groups showed reductions in peak compression from pre-to-post: on average the control group improved by 11.2%, the EMG group by 16.7%, and the acceleration group by 25.3%. The results revealed an interaction between the improvement and the group (p=0.023), and a difference between the improvement in the control group and that in the verbal acceleration feedback group (p<0.01). These reductions in lumbosacral compression persisted after a 7-day interval without feedback training, suggesting that this approach could provide sustained risk-reduction during manual materials handling.

The Son-Rise Program intervention for autism: an investigation into family experiences.

BACKGROUND: Despite the increasing involvement of parents as therapists in interventions for their children with autism, research to date has focused almost exclusively on the outcome for the child, and little is known about the effects of involvement on the whole family. This is true even of highly intensive home-based approaches such as the Son-Rise Program (SRP), the focus of the present paper. A longitudinal questionnaire-based study is reported which investigated a number of potential positive and negative effects for the family, how these changed over time, and their relation to child characteristics and patterns of intervention implementation. METHODS: Questionnaires examining family demographics, patterns of intervention use and perceived family effects were distributed three times over the course of a year to families who had attended an initial training course in the use of the SRP. RESULTS: The results indicated that, although involvement led to more drawbacks than benefits for the families over time, family stress levels did not rise in all cases. Few relationships were found between family effects and patterns of intervention use, although there was a strong connection with parental perceptions of intervention efficacy. CONCLUSIONS: The findings of the present study emphasize the need for those supporting families using home-based interventions to consider the needs of the whole family. This may be especially important if there are periods during which the family find the intervention to be less effective. Families embarking on such intensive approaches may also benefit from considering ways in which any disruption to family life can be minimized.

Multimodality radiotherapy and androgen ablation in the treatment of clinically localized prostate cancer: early results in high risk patients.

In patients presenting with clinically localized prostate cancer, the risk of biochemical failure increases significantly with higher Gleason scores, prostate specific antigen (PSA) levels, and clinical stages. Current surgical and radiotherapeutic approaches appear to offer limited success in patients with highly adverse prognostic factors. In an attempt to improve on these outcomes, we have combined external beam radiotherapy (EBRT) with a brachytherapy (BT) boost and neo adjuvant and adjuvant androgen ablation in a population at significant risk of biochemical failure. Here we present early biochemical progression data for this approach. From October 1997 to July 1999, 72 men with a serum PSA >or=10 ng/ml or Gleason score >or=7 or clinical stage >or=T2c (AJC/UICC 1992) underwent EBRT followed by palladium-103 BT. All patients underwent 8 months of combined androgen ablation with leuprolide and an oral antiandrogen beginning 3 months prior to initiation of EBRT. Patients were followed by PSA and digital rectal examination (DRE) at 3-month intervals and a chart review on all patients was carried out during July 2001. To allow comparisons to contemporary literature, Kaplan-Meier survival curves were generated utilizing three alternate definitions of biochemical recurrence: PSA >0.2 ng/ml, PSA >1.0 ng/ml, and the American Society for Therapeutic Radiology and Oncology (ASTRO) consensus definition of three consecutive rising PSAs. Our results indicate that when PSA >0.2 ng/ml was used to define biochemical progression, 88% (95% CI 80-97) of patients remained free of disease at 24 months. When PSA >1.0 ng/ml was used, 97% (CI 92-100) of patients remained disease free at 24 months. ASTRO criteria yielded 90% (CI 82-98) recurrence-free survival at 24 months. In conclusion, this very early report indicates that in patients who are at increased risk of biochemical failure, EBRT with a BT boost in conjunction with short-term androgen ablation offers potentially superior biochemical disease-free survival to contemporary alternative approaches in the literature. Clearly, longer follow-up is required to confirm the durability of this approach.

Head injury and alexithymia: implications for family practice care.

BACKGROUND: Alexithymia, a deficit in emotional information processing, and a history of head injury have both been found to be related to high rates of psychosomatic illness, substance abuse, depression, and utilization of primary care services. To date, no study has examined the potential comorbidity of alexithymia and head injury in a family practice setting, a necessary step in evaluating the aetiologic role of head injury in the development of alexithymia. The goals of this study are to establish prevalence of head injury and alexithymia in a family practice setting and to evaluate the relation, if present, between the two. METHODS: Patients (n =135) of a family practice residency facility were screened using the Traumatic Brain Injury Questionnaire and the Toronto Alexithymia Scale-20. RESULTS: Forty-nine per cent of the participants reported a history of head injury and 18% were alexithymic. Those with a history of head injury had significantly higher scores of alexithymia. Chi-square analysis indicated a relation between head injury and alexithymia. CONCLUSIONS: The high rates of self-reported history of head injury in family practice settings, particularly in the context of alexithymia, may adversely affect a physician's ability to care for these patients. Increasing physicians' awareness of head injury and the potential mediating role of alexithymia in medical and psychological illness may facilitate effective diagnosis and patient-physician communication.

Correcting out-of-plane errors in two-dimensional imaging using nonimage-related information.

Two-dimensional imaging with a single camera assumes that the motion occurs in a calibrated plane perpendicular to the camera axis. It is well known that kinematic errors result if the object fails to remain in this plane and that if both the distance to the calibration plane from the camera and the distance out-of-plane are known, an analytical correction for the out-of-plane error can be made. Less well appreciated is that out-of-plane distance can frequently be acquired from other, nonimage-related information. In the two examples given, the mediolateral center of pressure coordinate of the foot measured from a force plate and the measured landing point of a shot put throw were used. In both cases, the resulting out-of-plane correction improved the accuracy of the 2-D kinematic data dramatically. These examples also demonstrate that the use of nonimage-related data can increase the accuracy of kinematic data without an increase in the complexity of the experiment.

A finite element stress analysis of the transmandibular implant system.

PURPOSE: The aim of this work was a simulation of the onset of osseointegration of the transmandibular implant system in the mandible. This was achieved by imposing joint elements at the implant-bone interface. MATERIALS AND METHODS: The geometric model was derived from the anatomic measurement of several mandibles by tomographic scanning. The spring constants of the joint elements interposed at the implant-bone interface were varied between 1 N/mm and 10(9) N/mm to represent the conditions leading to osseointegration. RESULTS: Increasing the value of the spring constant at the joint elements gradually increases the effective stress in an increasing volume of crestal cortical bone. Additionally, a larger volume of crestal cortical bone assumes a higher stress value as the simulation proceeds. CONCLUSION: This work indicates that considerable changes in stress magnitude and distribution occur in the crestal cortical bone margins as osseointegration is simulated, which may be the necessary stimulus for bone remodeling.

PRMT1 is the predominant type I protein arginine methyltransferase in mammalian cells.

Type I protein arginine methyltransferases catalyze the formation of asymmetric omega-N(G),N(G)-dimethylarginine residues by transferring methyl groups from S-adenosyl-L-methionine to guanidino groups of arginine residues in a variety of eucaryotic proteins. The predominant type I enzyme activity is found in mammalian cells as a high molecular weight complex (300-400 kDa). In a previous study, this protein arginine methyltransferase activity was identified as an additional activity of 10-formyltetrahydrofolate dehydrogenase (FDH) protein. However, immunodepletion of FDH activity in RAT1 cells and in murine tissue extracts with antibody to FDH does not diminish type I methyltransferase activity toward the methyl-accepting substrates glutathione S-transferase fibrillarin glycine arginine domain fusion protein or heterogeneous nuclear ribonucleoprotein A1. Similarly, immunodepletion with anti-FDH antibody does not remove the endogenous methylating activity for hypomethylated proteins present in extracts from adenosine dialdehyde-treated RAT1 cells. In contrast, anti-PRMT1 antibody can remove PRMT1 activity from RAT1 extracts, murine tissue extracts, and purified rat liver FDH preparations. Tissue extracts from FDH(+/+), FDH(+/-), and FDH(-/-) mice have similar protein arginine methyltransferase activities but high, intermediate, and undetectable FDH activities, respectively. Recombinant glutathione S-transferase-PRMT1, but not purified FDH, can be cross-linked to the methyl-donor substrate S-adenosyl-L-methionine. We conclude that PRMT1 contributes the major type I protein arginine methyltransferase enzyme activity present in mammalian cells and tissues.

Elevated levels of Ca(II) modulate the activity and inhibition of serine proteases: implication in the mechanism of apoptosis.

Elevated levels of intracellular Ca(II) are a prominent feature of apoptosis, a natural form of cell death involved in many physiological and pathological processes. Serine proteases play crucial roles in apoptosis and have been implicated in the genomic DNA degradation and the massive protein degradation that occur during apoptosis. In this study, the effects of the elevated level of Ca(II) on the activity and inhibition of serine proteases were examined by spectrophotometric methods. The effects of the elevated levels of Ca(II), Mg(II), K(I), and Na(I) on the activity and inactivation of three representative members of serine proteases were determined. The level of serine protease activity in CEM-C7-14 leukemic cells was also evaluated in the presence and absence of dexamethasone-induced apoptosis, and also in the presence of A23187, a Ca(II)-ionophore. Among the four metal-ions studied, only Ca(II) was found to significantly enhance the activity of mammalian serine proteases. Ca(II) was also found to significantly protect the enzymes from inhibition, while the other three metal-ions showed no significant effect on the inactivation of the enzymes. Compared to the control sample, the enzymic activity was found to be higher during apoptosis, and in the presence of the Ca(II)-ionophore. Results of this study indicate that Ca(II) can significantly enhance the catalytic efficiency of serine proteases during apoptosis.

Identification of the RNA binding domain of T4 RegA protein by structure-based mutagenesis.

The T4 translational repressor RegA protein folds into two structural domains, as revealed by the crystal structure (Kang, C.-H. , Chan, R., Berger, I., Lockshin, C., Green, L., Gold, L., and Rich, A. (1995) Science 268, 1170-1173). Domain I of the RegA protein contains a four-stranded beta-sheet and two alpha-helices. Domain II contains a four-stranded beta-sheet and an unusual 3/10 helix. Since beta-sheet residues play a role in a number of protein-RNA interactions, one or both of the beta-sheet regions in RegA protein may be involved in RNA binding. To test this possibility, mutagenesis of residues on both beta-sheets was performed, and the effects on the RNA binding affinities of RegA protein were measured. Additional sites for mutagenesis were selected from molecular modeling of RegA protein. The RNA binding affinities of three purified mutant RegA proteins were evaluated by fluorescence quenching equilibrium binding assays. The activities of the remainder of the mutant proteins were evaluated by quantitative RNA gel mobility shift assays using lysed cell supernatants. The results of this mutagenesis study ruled out the participation of beta-sheet residues. Instead, the RNA binding site was found to be a surface pocket formed by residues on two loops and an alpha-helix. Thus, RegA protein appears to use a unique structural motif in binding RNA, which may be related to its unusual RNA recognition properties.

Determination of molecular masses of proteins in solution: Implementation of an HPLC size exclusion chromatography and laser light scattering service in a core laboratory.

Size exclusion chromatography (SEC) coupled with "on-line" laser light scattering (LS), refractive index (RI), and ultraviolet (UV) detection provides an elegant approach to determining the molecular weights of proteins and their complexes in solution. SEC serves solely as a fractionation step to minimize the ambiguity that otherwise can result from the fact that light scattering provides the weight-average molecular weight (MW) of all species in solution. Our goal is to establish realistic expectations for MW determination using LS coupled with SEC, define sample requirements, and identify possible limitations of SEC/LS analysis. Analyses of 14 protein standards that range from 12 to 475 kd suggest that the molecular weights of native proteins may be determined in a single SEC/LS experiment with an accuracy of +/-5%. The MW determination depends only on the downstream LS and RI detectors, and it is independent of elution position. Unusual elution because of nonglobular shape or interaction with the SEC support has no impact on the MW determination by SEC/LS. With the instrument configuration that was used, the optimal amount of protein needed for SEC/LS is about 50 g for proteins with molecular weight greater than 40 kd. However, analyses of ovalbumin and transferrin demonstrate that even 10 g is sufficient to determine the MW with an error of less than +/-6%. Although SEC/LS has some limitations, such as proteins that contain chromophores whose absorption spectrum overlaps that of the emission spectrum of the laser, it represents a fast and robust approach to determining MW and to monitoring protein oligomerization in solution.

Uptake and internalization of exogenous apolipoprotein E3 by cultured human central nervous system neurons.

Apolipoprotein E (apoE) has been confirmed as a risk factor for late-onset Alzheimer's disease (AD) and is associated with neurofibrillary tangles and senile plaques, the microscopic pathological characteristics of AD. There has been no direct evidence that human central nervous system neurons can take up and internalize exogenous apoE, which may be important in order for apoE to be involved in the development of the disease. This paper demonstrates by immunohistochemistry and confocal microscopy that cultured human brain neurons can take up and internalize exogenous recombinant human apoE3. We confirm that neurons express the low-density lipoprotein receptor-related protein (LRP) but do not express the low-density lipoprotein receptor. We also demonstrate that the LRP mediates the neuronal uptake of apoE.

Identification of protein-arginine N-methyltransferase as 10-formyltetrahydrofolate dehydrogenase.

S-Adenosylmethionine:protein-arginine N-methyltransferase (EC 2.1.1. 23; protein methylase I) transfers the methyl group of S-adenosyl-L-methionine to an arginine residue of a protein substrate. The homogeneous liver protein methylase I was subjected to tryptic digestion followed by reverse phase high performance liquid chromatography (HPLC) separation and either "on-line" mass spectrometric fragmentation or "off-line" Edman sequencing of selected fractions. Data base searching of both the mass spectrometric and Edman sequencing data from several peptides identified the protein methylase as 10-formyltetrahydrofolate dehydrogenase (EC 1.5.1.6; Cook, R. J., Lloyd, R. S., and Wagner, C. (1991) J. Biol. Chem. 266, 4965-4973; Swiss accession number). This identification was confirmed by comparative HPLC tryptic peptide mapping and affinity chromatography of the methylase on the 5-formyltetrahydrofolate-Sepharose affinity gel used to purify the dehydrogenase. The purified rat liver methylase had approximately 33% of the 10-formyltetrahydrofolate dehydrogenase and 36% of the aldehyde dehydrogenase activity as compared with the recombinant dehydrogenase, which also had protein methylase I activity. Polyclonal antibodies against recombinant dehydrogenase reacted with protein methylase I purified either by polyacrylamide gel electrophoresis or 5-formyltetrahydrofolate affinity chromatography. In each instance there was only a single immunoreactive band at a molecular weight of approximately 106,000. Together, these results confirm the co-identity of protein-arginine methyltransferase and 10-formyltetrahydrofolate dehydrogenase.

Use of liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS) for routine identification of enzymatically digested proteins separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis.

Automated liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS) analysis of >100 tryptic digests carried out on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) separated, Coomassie Blue-stained proteins that were prepared by >50 different laboratories demonstrates that a commercial electrospray/quadrupole ion trap mass spectrometer and the tandem mass correlation algorithm developed by Eng et al. (Am. Soc. Mass Spectrom. 1994, 5, 976-989) provide an extremely robust and facile approach to routine protein identification. By requiring a minimum of two significant matches to peptides that would be predicted to be produced by the protease that was used, low pmol levels of proteins can be identified with high confidence while minimizing the probability of identifying the protease itself and/or the ubiquitous contaminant, keratin. Hence, in only 7% of the digests analyzed was keratin identified and in only 5% of the digests analyzed was the protease itself identified. In contrast, 58% of the analyzed samples were identified and, in many instances, multiple proteins were identified in the same sample. Although the median amount of digest analyzed was 6.1 pmol, the limit of sensitivity (as the instrument is configured with a flow rate of 4 microL/min) appears to be at the 500 fmol level. Since one of the primary reasons for not identifying a sample is that its sequence is not yet in the database searched, the utility of an LC MS/MS approach to protein identification will certainly increase in the future as the sequences of more genomes are completed.

Design of prosthetic cantilever bridgework supported by osseointegrated implants using the finite element method.

OBJECTIVES: The aim of the present work was to establish a design procedure for fixed metal prostheses supported by osseointegrated implants in order to prevent permanent deformation and hence failure following loading. Previously, the cantilever cross-sectional shape in the buccal lingual plane has been based on clinical experience and subjectivity. METHODS: This work has relied on the use of linear elastic finite element analysis in order to generate a maximum effective stress at which permanent deformation commences on loading. A number of different cross-sectional shapes were investigated, both of conventional design as well as new innovative possibilities. Both straight and curved cantilever beams 26 mm long were examined. RESULTS: The design failure chosen was based on a von Mises plastic collapse principle by comparing the calculated effective stresses with the yield stress of the metal in simple tension. It was found that the "L" shaped design was more rigid than other designs for a given mass, while a framework based on an open "I" section offers good possibilities particularly when used as curved shapes. SIGNIFICANCE: Assuming a failure criterion based on the von Mises principle, then "L" shaped Co/Cr or stainless steel frameworks, typically 26 mm of cantilever span, undergo permanent deformation at end loadings between 130 and 140 N depending on section curvature. Since it is known biting loads can exceed these values, good design is critical if such failures are to be avoided.